Misreading, a fundamental aspect of the mechanism of action of several aminoglycosides

Lando, Danielle; Cousin, Marie-Anne; Garilhe, M. Privat de

doi:10.1021/bi00746a035

Cited by 28 publications

(11 citation statements)

References 21 publications

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“…2B). In contrast, ethanol did not show cooperative synergy with chloramphenicol, which inhibits peptidyl transfer in the ribosome but does not decrease translational accuracy (37), suggesting that ethanol does not impede the peptidyl transfer reaction. These results strongly indicate that sublethal ethanol stress induces physiologically damaging levels of translational misreading, perhaps compromising the conformation of the ribosomal decoding site known to be targeted by streptomycin (38,39).…”

Section: Resultsmentioning

confidence: 76%

Correcting direct effects of ethanol on translation and transcription machinery confers ethanol tolerance in bacteria

Haft

Keating

Schwaegler

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

130

121

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The molecular mechanisms of ethanol toxicity and tolerance in bacteria, although important for biotechnology and bioenergy applications, remain incompletely understood. Genetic studies have identified potential cellular targets for ethanol and have revealed multiple mechanisms of tolerance, but it remains difficult to separate the direct and indirect effects of ethanol. We used adaptive evolution to generate spontaneous ethanol-tolerant strains of Escherichia coli, and then characterized mechanisms of toxicity and resistance using genome-scale DNAseq, RNAseq, and ribosome profiling coupled with specific assays of ribosome and RNA polymerase function. Evolved alleles of metJ, rho, and rpsQ recapitulated most of the observed ethanol tolerance, implicating translation and transcription as key processes affected by ethanol. Ethanol induced miscoding errors during protein synthesis, from which the evolved rpsQ allele protected cells by increasing ribosome accuracy. Ribosome profiling and RNAseq analyses established that ethanol negatively affects transcriptional and translational processivity. Ethanol-stressed cells exhibited ribosomal stalling at internal AUG codons, which may be ameliorated by the adaptive inactivation of the MetJ repressor of methionine biosynthesis genes. Ethanol also caused aberrant intragenic transcription termination for mRNAs with low ribosome density, which was reduced in a strain with the adaptive rho mutation. Furthermore, ethanol inhibited transcript elongation by RNA polymerase in vitro. We propose that ethanol-induced inhibition and uncoupling of mRNA and protein synthesis through direct effects on ribosomes and RNA polymerase conformations are major contributors to ethanol toxicity in E. coli, and that adaptive mutations in metJ, rho, and rpsQ help protect these central dogma processes in the presence of ethanol.

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Section: Resultsmentioning

confidence: 76%

Correcting direct effects of ethanol on translation and transcription machinery confers ethanol tolerance in bacteria

Haft

Keating

Schwaegler

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

130

121

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“…b) Streptomycin and dihydrostreptomycin induce a lower level of misreading [30] and are more potent inhibitors of the initiation of translation than paromomycin [10,31,32]. Since their mechanism of action is different from that of paromomycin it is not surprising that they should have a different binding site.…”

Section: Discussionmentioning

confidence: 99%

Paromomycin and Dihydrostreptomycin Binding to Escherichia coli Ribosomes

Lando¹,

Cousin²,

Ojasoo³

et al. 1976

European Journal of Biochemistry

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Paromomycin binds specifically to a single type of binding site on the 70‐S streptomycin‐sensitive Escherichia coli ribosome. This site is different from that of dihydrostreptomycin since paromomycin binds to streptomycin‐resistant ribosomes and since dihydrostreptomycin does not compete for paromomycin binding. Paromomycin binding, unlike dihydrostreptomycin binding, is independent of changes in ribosome concentration but influenced by magnesium ion concentration. Moreover, paromomycin does not bind to the 30‐S subunit of the streptomycin‐sensitive ribosome, except in the presence of dihydrostreptomycin, which probably induces the conformational changes necessary for a paromomycin binding site. This induction does not occur with streptomycin‐resistant ribosomes. Neither antibiotic binds to the 50‐S subunit. In general, binding of the one antibiotic increases the number of sites available for binding of the other. Both antibiotics hibit marked non‐specific binding at high antibiotic/ribosome ratios. Competition studies have enabled the classification of other aminoglycosides according to their ability to compete for the paromomycin and dihydrostreptomycin binding sites. Derivatives structurally related to paromomycin compete for its binding, the degree of competition being related to antibacterial activity, but do not compete for dihydrostreptomycin binding; they, on the contrary, increase the number of dihydrostreptomycin binding sites. Neither gentamicin nor kanamycin derivatives, which induce a high level of misreading, nor kasugamycin and spectinomycin, which do not induce misreading, compete for paromomycin or dihydrostreptomycin binding sites. Other sites may be involved in the binding of these aminoglycosides and in inducing misreading.

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“…In order to obtain more information a b h i the binding site [5] of this class of molecules, competition experiments were performed using [14C]acetyltobramycin as a marker. Molecules almost identical in structure (tobramycin, kanamycin A, B and C, gentamicin CIA), others somewhat related in structure (streptomycin, neomycin, dihydrosisomycin) and one not at all related in structure (erythromycin) were chosen as possible competitors.…”

Section: Competition Hpevimentsmentioning

confidence: 99%

“…Their action can be explained as either an inhibition of protein biosynthesis itself or a misreading of the genetic code. Experimental findings correlating the structures of these drugs to either type of perturbation [3] have been published, but apart from streptomycin [4] and paromomycin [5], which have been studied in terms of binding experiments, little is known about the aminoglycoside antibiotics in which both C-4 and C-6 hydroxyl groups of the deoxystreptamine moiety are substituted by aminosugar residues. However, a recent report [6] described the binding [3H]-kanamycin A to washed ribosomes and its subunits.…”

mentioning

confidence: 99%

Mechanism of Action of Aminoglycoside Antibiotics. Binding Studies of Tobramycin and Its 6'-N-Acetyl Derivative to the Bacterial Ribosome and Its Subunits

et al. 1979

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6′‐N‐[14C]Acetyl‐tobramycin and [3H]tobramycin were synthesized and their binding to Escherichia coli ribosomes and ribosomal subunits studied using equilibrium dialysis. The 70‐S ribosome, as well as its 50‐S and 30‐S subunits, bound tightly to 6′‐N‐acetyl‐tobramycin. The binding of [3H]tobramycin to ribosomes was quite different. The 70‐S ribosome was observed to possess several classes of binding sites; of these, one was determined to be of higher affinity and lower capacity, the 6′‐N‐[14C]acetyl‐tobramycin site. The isotopic dilution method was used to define the specificity of the interaction. The selective binding of 6′‐N‐[14C]acetyl‐tobramycin was highly reversible by tobramycin, kanamycins A, B, C and neomycin, but not by streptomycin or erythromycin. Gentamicin C1a was a poor inhibitor. This suggested that either the kanosamin or garosamin rings might be determinant in the binding of these molecules, as well as the 6′‐amino group.

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Misreading, a fundamental aspect of the mechanism of action of several aminoglycosides

Cited by 28 publications

References 21 publications

Correcting direct effects of ethanol on translation and transcription machinery confers ethanol tolerance in bacteria

Correcting direct effects of ethanol on translation and transcription machinery confers ethanol tolerance in bacteria

Paromomycin and Dihydrostreptomycin Binding to Escherichia coli Ribosomes

Mechanism of Action of Aminoglycoside Antibiotics. Binding Studies of Tobramycin and Its 6'-N-Acetyl Derivative to the Bacterial Ribosome and Its Subunits

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